Basophil CD63 expression assay on highly sensitized atopic donor leucocytes-a useful method in chronic autoimmune urticaria

Abstract

Our new technical observation demonstrates that basophils of highly sensitized atopic donors can be successfully used without priming with IL-3 for the in-vitro flow cytofluorimetric diagnosis of CU. With this investigation the characterization of the autoimmune origin of CU is based on an objective in vitro technique.

Clinical and Laboratory Examinations in the Subgroups of Chronic Urticaria

“…The CD63 expression on the surface of donor basophils was determined as described earlier [20]. Briefly, after incubation with chronic urticaria sera, the cells were labelled with goat anti-human IgE FITC and R-phycoerythrin conjugated anti-human CD63 monoclonal antibody (both from Caltag, Burlingame, Calif., USA) and the percentage of IgE and CD63 double-positive basophils was measured on a Becton Dickinson FACSCalibur flow cytometer.…”

“…One of them, the gold standard, is the basophil histamine release (HR) test [18]. In recent studies, a new method, the modified CD63 expression assay, was found to be a reliable method for the diagnosis of AIU [19,20,21,22]. …”

Serum induced CD63 and CD203c activation tests in chronic urticaria

“…After ROC curve analysis, we concluded that CD203c did not have sufficient diagnostic power. Other authors, who tested CD63 [11][12][13][14][15] or CD203c expression [17] in CU patients, found significant correlation between ASST and both basophil activation tests positivity. The discrepancy between CD63 and CD203c correlations with ASST in our study can reflect either different mechanism of CD63 and CD203c molecules upregulation or differences in methodology of tests.…”

“…Recently, the ability of CU patient's serum to evoke expression of CD63 and CD203c on donor human basophils assessed by flow cytometry has been tested. There are a few published studies assessing serum-induced CD63 expression measurement as a diagnostic tool in CAU [11][12][13][14][15][16] and only 2 studies done by measured CD203c expression [16,17].…”

Chronic urticaria and thyroid autoimmunity markers

“…The basophil histamine release assay (HRA) is currently the "gold standard" for detecting functional antibodies in the serum of patients with CU [5][6][7]. Improved screening tests have been sought in the last decade; for instance, flow cytometry used to determine the ability of a CU patient's serum to evoke an expression of CD63 and CD203c on donor basophils is showing some promise [8][9][10][11][12][13].…”